Isolation and Characterization of Cardiac Mesenchymal Stromal Cells from Endomyocardial Bioptic Samples of Arrhythmogenic Cardiomyopathy Patients

Abstract

A normal adult heart is composed of several different cell types, among which cardiac mesenchymal stromal cells represent an abundant population. The isolation of these cells offers the possibility of studying their involvement in cardiac diseases, and, in addition, provides a useful primary cell model to investigate biological mechanisms.Here, the method for the isolation of C-MSC from arrhythmogenic cardiomyopathy patients' bioptic samples is described. The endomyocardial biopsy sampling is guided in the right ventricular areas adjacent to the scar visualized by electro-anatomical mapping. The digestion of the biopsies in collagenase and their plating on a plastic dish in culture medium to allow C-MSC growth is described. The isolated cells can be expanded in culture for several passages. To confirm their mesenchymal phenotype, the description of immuno-phenotypical characterization is provided. C-MSC are able to differentiate into several cell types like adipocytes, chondrocytes, and osteoblasts: in the context of ACM, characterized by adipocyte deposits in patients' hearts, the protocols for the adipogenic differentiation of C-MSC and the characterization of lipid droplet accumulation are described.

Fibrosis in Arrhythmogenic Cardiomyopathy: The Phantom Thread in the Fibro-Adipose Tissue

“…In the injured mouse heart, as during myocardial infarction, C-MSC resident population (not recruited from the bone marrow) express stem cell and fibroblast markers like collagen type I and DDR2, suggesting their involvement in scar formation (Carlson et al, 2011). C-MSC have been involved as major player of ACM adipogenesis (Sommariva et al, 2016;Pilato et al, 2018). A C-MSC population isolated based on PDGFRα and Sca1 could be responsible for fibrofatty scar formation in ACM patients.…”

Differences in Mitochondrial Membrane Potential Identify Distinct Populations of Human Cardiac Mesenchymal Progenitor Cells

“…TMRM-high and TMRM-low populations were plated at a concentration of 30,000 cells/cm 2 and cultured in adipogenic conditions for 1 week [ 58 ]. The adipogenic medium consists of IMDM supplemented with 10% FBS (Sigma-Aldrich, Milan, Italy), 0.5 mmol/L 3-isobutyl-1-methylxanthine (Sigma-AldrichMilan, Italy), 1 µmol/L hydrocortisone (Sigma-Aldrich, Milan, Italy), 0.1 mmol/L indomethacin (Sigma-Aldrich, Milan, Italy), 10,000 U/mL Penicillin (Thermo Fisher Scientific, Milan, Italy), 10,000 µg/mL Streptomycin (Thermo Fisher Scientific, Milan, Italy) and 20 mmol/L L-Glutamine (Sigma-Aldrich, Milan, Italy).…”

Mesenchymal stromal cell‐derived extracellular vesicles as cell‐free biologics for the ex vivo expansion of hematopoietic stem cells

“…MSCs were first isolated from the BM (Friedenstein et al, 1968), but recent studies have revealed that these cells can be cultured from various other tissues such as adipose (Estes et al, 2010), heart (Pilato et al, 2018), Wharton's jelly (Chao et al, 2008), peripheral blood (He et al, 2007), dental pulp (Jo et al, 2007), cord blood (Oh et al, 2008), menstrual blood (Patel et al, 2008), and chorionic villi (Yang et al, 2013). MSCs are characterized by the presence of surface markers such as CD105, CD73, CD90 and the absence of CD45, CD34, CD14, CD11b, CD79a, or CD19 and HLA‐DR as well as the ability to differentiate into osteoblasts, chondrocytes, and adipocytes (Dominici et al, 2006).…”